摘要
OBJECTIVE: Celastrol has been established as a nuclear factor-κB(NF-κB) activation inhibitor; however, the exact mechanism behind this action is still unknown. Using text-mining technology, the authors predicted that int erleukin-1 receptor-associated kinases(IRA Ks) are potential celastrol targets, and hypothesized that targeting IRAKs might be one way that celastrol inhibits NF-κB. This is because IRAKs are key molecules for some crucial pathways to activate NF-κB(e.g., the inter leukin-1 receptor(IL-1R)/Toll- like receptor(TLR) superfamily).METHODS: The human hepatocellular cell line(Hep G2) treated with palmitic acid(PA) was used as a model for stimulating TLR4/NF-κB activation, in order to observe the potential effects of celastrol in IRAK regulation and NF-κB inhibition. The transfection of small interfering RNA was used for down-regulating TLR4, IRAK1 and IRAK4, and the Western blot method was used to detect changes in the protein expressions.RESULTS: The results showed that celastrol could effectively inhibit PA-caused TLR4-dependent NF-κB activation in the Hep G 2 cells; PA also activated IRAKs, which were inhibited by celastrol. Knocking down IRAKs abolished PA-caused NF-κB activation.CONCLUSION: The results for the first time show that targeting IRAKs is one way in which celastrol inhibits NF-κB activation.
OBJECTIVE: Celastrol has been established as a nuclear factor-κB(NF-κB) activation inhibitor; however, the exact mechanism behind this action is still unknown. Using text-mining technology, the authors predicted that int erleukin-1 receptor-associated kinases(IRA Ks) are potential celastrol targets, and hypothesized that targeting IRAKs might be one way that celastrol inhibits NF-κB. This is because IRAKs are key molecules for some crucial pathways to activate NF-κB(e.g., the inter leukin-1 receptor(IL-1R)/Toll- like receptor(TLR) superfamily).METHODS: The human hepatocellular cell line(Hep G2) treated with palmitic acid(PA) was used as a model for stimulating TLR4/NF-κB activation, in order to observe the potential effects of celastrol in IRAK regulation and NF-κB inhibition. The transfection of small interfering RNA was used for down-regulating TLR4, IRAK1 and IRAK4, and the Western blot method was used to detect changes in the protein expressions.RESULTS: The results showed that celastrol could effectively inhibit PA-caused TLR4-dependent NF-κB activation in the Hep G 2 cells; PA also activated IRAKs, which were inhibited by celastrol. Knocking down IRAKs abolished PA-caused NF-κB activation.CONCLUSION: The results for the first time show that targeting IRAKs is one way in which celastrol inhibits NF-κB activation.
基金
the Shanghai Gongli Hospital Youth Project (No. 2014GLQN16 for YS and No. 2012GLQN09 for XZ)
the Shanghai Pudong District Science and Technology Innovation Project (No. PKJ2013-Y03 for YW)
the Shanghai Pudong Youth Talent Project in Medicine (No. PWRq2013-11 for FC)
the Shanghai Yang Fan Project (No. 15YF1410800 for FC)
the International Science & Technology Cooperation Project of China (Grant 2011DFB30010 for GU)
the National Natural Science Foundation of China (No. 81102349 for BP and No. 81400793 for YW)
the Shanghai Excellent Academic Leader in Medicine (No. XBR2011054 for DZ)
the Shanghai Traditional Chinese Medicine Content Construction Innovation Project (No. ZY3-CCCX-3-7001 for DZ)